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[Evaluation of Toll-Like Receptor Gene Expressions in Encephalitozoon intestinalis Infection]. [评估肠道脑线虫感染中的 Toll-Like 受体基因表达]。
IF 1 4区 医学 Q4 MICROBIOLOGY Pub Date : 2024-04-01 DOI: 10.5578/mb.202498201
Erdi Uzun, Ülfet Çetinkaya
<p><p>Microsporidia are obligate intracellular pathogens that can infect many vertebrate and invertebrate hosts. While the Microsporidia phylum was defined as protozoa until the 1990s, it has been associated with fungi in line with the data obtained as a result of phylogenetic and molecular analyzes in recent years. Although approximately 200 genera and 1400 Microsporidia species related to these genera have been reported to date, only 14 species are known to cause infection in humans. Encephalitozoon intestinalis is one of the most frequently detected species in humans and causes serious clinical conditions in immunosuppressed individuals. Little information is available about the immunology of this infection. This study was aimed to investigate the changes in Toll-Like receptor (TLR) gene expressions in Madin-Darby canine kidney (MDCK) cells treated with E.intestinalis spores. Three groups were formed in the study. In the first group, only the medium prepared for E.intestinalis was added to the MDCK cells. In the second group, 108 live spores waiting at +4 °C were added. In the third group, 108 heat-inactivated spores were added. All three groups were incubated at 37ºC with 5% CO2 . RNA isolation and cDNA synthesis were performed from samples taken from these groups at the 1st, 3rd, 6th, 12th and 24th hours. Expression of TLR1-10 genes from the obtained cDNAs was evaluated by real-time polymerase chain reaction (Rt-PCR). GAPDH and ACTB genes were used as housekeeping genes in the study. Target genes were normalized by taking the average of these two genes and statistical analysis was performed by applying the 2-ΔΔCt formula. Genes detected above the threshold value (threshold 1) were considered to have increased expression. Genes detected below the threshold value were considered to have decreased expression. The growth of the live and inactive spores were followed simultaneously with the experimental groups. Approximately two weeks after the start of the culture, it was observed that E.intestinalis grew in the culture with live spore, but did not grow in the culture with inactivated spores. No statistically significant change was observed in gene expressions in the inactivated spore group. In the live spore group, a significant increase was seen in the expression of only two genes. These genes were TLR3 and TLR4. It was observed that there was a significant increase in TLR3 gene expression at the first hour (1.6-fold of control group) but the expression level started to decrease at the third hour (1.4-fold of control group) and returned to the control level at the sixth hour. It was observed that TLR4 gene expression continued parallel to the control until the 24th hour and increased significantly (2.1-fold of control group) at the 24th hour. In conclusion, this study is the f irst report in which the changes in ten different TLR gene expressions were evaluated at different times in MDCK cells stimulated with E.intestinalis and the change in T
小孢子虫是一种细胞内强制性病原体,可感染许多脊椎动物和无脊椎动物宿主。虽然在 20 世纪 90 年代之前,小孢子虫门一直被定义为原生动物,但近年来,根据系统发育和分子分析获得的数据,小孢子虫门已与真菌联系在一起。尽管迄今已报道了与这些属相关的约 200 个属和 1400 个小孢子虫种,但已知只有 14 个种可导致人类感染。肠脑线虫是人类最常检测到的种类之一,会导致免疫抑制人群出现严重的临床症状。目前有关这种感染的免疫学信息很少。本研究旨在调查经肠孢子虫处理的马丁达比犬肾细胞(MDCK)中Toll-Like受体(TLR)基因表达的变化。研究分为三组。第一组,只向 MDCK 细胞中添加为肠杆菌属孢子制备的培养基。第二组,加入 108 个在 +4 °C下等待的活孢子。第三组加入 108 个热灭活孢子。所有三组均在 37ºC 和 5% CO2 条件下培养。在第 1、第 3、第 6、第 12 和第 24 小时从这些组别中提取样本进行 RNA 分离和 cDNA 合成。通过实时聚合酶链式反应(Rt-PCR)评估所获得 cDNA 中 TLR1-10 基因的表达。研究中使用 GAPDH 和 ACTB 基因作为管家基因。目标基因通过取这两个基因的平均值进行归一化,并应用 2-ΔΔCt 公式进行统计分析。检测到高于阈值(阈值 1)的基因被认为表达量增加。检测到的基因低于阈值,则认为表达量减少。实验组同时跟踪活孢子和非活性孢子的生长情况。培养开始约两周后,观察到肠杆菌在活孢子培养物中生长,但在失活孢子培养物中没有生长。灭活孢子组的基因表达没有明显的统计学变化。在活孢子组中,只有两个基因的表达量有明显增加。这两个基因是 TLR3 和 TLR4。据观察,TLR3 基因的表达量在第一小时明显增加(是对照组的 1.6 倍),但在第三小时开始下降(是对照组的 1.4 倍),在第六小时恢复到对照组的水平。据观察,TLR4 基因的表达一直与对照组平行,直到第 24 小时,并在第 24 小时显著增加(对照组的 2.1 倍)。总之,本研究首次报道了在肠杆菌刺激的 MDCK 细胞中,十种不同 TLR 基因表达在不同时间的变化以及 TLR3 基因表达的变化。
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引用次数: 0
[Molecular Analysis of Antibiotic Resistance in CarbapenemResistant Pseudomonas aeruginosa Isolates Isolated from Clinical Samples]. [从临床样本中分离的耐碳青霉烯类铜绿假单胞菌抗生素耐药性的分子分析]。
IF 1 4区 医学 Q4 MICROBIOLOGY Pub Date : 2024-04-01 DOI: 10.5578/mb.202498200
Gülşah Altan, Erva Rakici, Osman Birol Özgümüş
<p><p>Pseudomonas aeruginosa is an opportunistic pathogen that causes increased morbidity and mortality in risky patient groups. Nowadays, carbapenem resistance has become a threat and resistance genes are spreading among species through mobile genetic elements. The dissemination of carbapenemases among P.aeruginosa is a serious public health concern due to its limited options for the treatment of bacterial infections. In this study, it was aimed to investigate the molecular epidemiology of 47 carbapenem resistant P.aeruginosa (CRPA) isolates derived from various clinical samples from the Central Laboratory Bacteriology Unit of Kocaeli University Research and Training Hospital between October 2021 and March 2023. The rates of resistance to the antibiotics, some carbapenemase and virulence genes, conjugative resistance plasmids, integron gene cassette contents and the clonal similarity of the isolates were investigated and then epidemiologically evaluated. In the study, identification of the bacterial isolates and their susceptibility to some antibiotics (imipenem, meropenem, aztreonam, amikacin, netilmicin, tobramycin, piperacillin, piperacillin/tazobactam, ceftazidime, cefepime, ciprofloxacin and levofloxacin) were determined by the VITEK® 2 Compact automated system. Metallo-beta-lactamase (MBL) production of the isolates was demonstrated by the imipenem/meropenem-EDTA (IMP/MEM-EDTA) combined disc method. Conjugation experiments were performed by the broth mating method. Alkali lysis method was used in plasmid DNA isolations. Co-transferred antibiotic resistances in transconjugants were detected by disc diffusion method. Carbapenemase genes (blaIMP , blaVIM , blaNDM , blaKPC and blaOXA-48 ), integron gene cassettes (class 1 and class 2) and virulence genes (lasR and rhlR) were screened by specific polymerase chain reactions (PCRs). Clonal relationships of the CRPA isolates were investigated by evaluating the DNA f ingerprintings obtained from the ERIC (enterobacterial repetitive intergenic consensus)-PCR assay. The highest resistance rate of the isolates were to levofloxacin, while the lowest resistance rates were observed against tobramycin, gentamicin and amikacin. MBL production was detected in 25 (53.2%) isolates. In conjugation experiments, 12 (25.5%) isolates were detected to harbour conjugative resistance plasmids. In 90% of the CRPA isolates, lasR and rhlR biofilm genes (encoding for the transcriptional activator protein) were detected by PCR. The blaVIM gene was detected in six (12.8%) isolates. The blaNDM gene was detected in five (10.6%) isolates and the blaOXA-48 gene was detected in three (6.4%) isolates. The blaKPC and blaIMP genes were not detected in CRPA isolates. It was determined that two (16.6%) of the isolates that carried the blaVIM gene, one (8.3%) carried the blaNDM gene and one (8.3%) carried the blaOXA-48 gene contained conjugative plasmids.In integron-specific PCRs, intI1 gene was positive in 39 (82.9%) isolates, while
铜绿假单胞菌是一种机会性病原体,会增加高危患者群体的发病率和死亡率。如今,铜绿假单胞菌对碳青霉烯类耐药已成为一种威胁,耐药基因正通过移动遗传因子在不同物种间传播。由于治疗细菌感染的选择有限,碳青霉烯酶在铜绿假单胞菌中的传播是一个严重的公共卫生问题。本研究旨在调查 2021 年 10 月至 2023 年 3 月期间科贾埃利大学研究与培训医院中央实验室细菌学组从各种临床样本中分离出的 47 例耐碳青霉烯类细菌(CRPA)的分子流行病学。对分离菌株的抗生素耐药率、部分碳青霉烯酶和毒力基因、共轭耐药质粒、整合子基因盒内容和克隆相似性进行了调查,然后进行了流行病学评估。在这项研究中,细菌分离物的鉴定及其对某些抗生素(亚胺培南、美罗培南、阿曲南、阿米卡星、奈替米星、妥布霉素、哌拉西林、哌拉西林/他唑巴坦、头孢他啶、头孢吡肟、环丙沙星和左氧氟沙星)的敏感性由 VITEK® 2 Compact 自动系统测定。亚胺培南/美罗培南-EDTA(IMP/MEM-EDTA)组合圆片法证明了分离菌株产生的金属-β-内酰胺酶(MBL)。肉汤配对法进行了共轭实验。质粒 DNA 分离采用碱裂解法。通过圆盘扩散法检测转染株的共转抗生素耐药性。通过特异性聚合酶链反应(PCR)筛选碳青霉烯酶基因(blaIMP、blaVIM、blaNDM、blaKPC 和 blaOXA-48)、整合子基因盒(1 类和 2 类)和毒力基因(lasR 和 rhlR)。通过评估从 ERIC(肠道细菌重复基因间共识)-PCR 检测中获得的 DNA 指纹,研究了 CRPA 分离物的克隆关系。分离株对左氧氟沙星的耐药率最高,而对妥布霉素、庆大霉素和阿米卡星的耐药率最低。在 25 个(53.2%)分离物中检测到了 MBL 的产生。在共轭实验中,检测到 12 个(25.5%)分离物携带共轭抗性质粒。在 90% 的 CRPA 分离物中,通过 PCR 检测到了 lasR 和 rhlR 生物膜基因(编码转录激活蛋白)。在 6 个(12.8%)分离株中检测到 blaVIM 基因。在 5 个分离株(10.6%)中检测到 blaNDM 基因,在 3 个分离株(6.4%)中检测到 blaOXA-48 基因。在 CRPA 分离物中未检测到 blaKPC 和 blaIMP 基因。在整合子特异性 PCR 中,39 个分离物(82.9%)的 intI1 基因呈阳性,24 个分离物(51%)检测到 1 类整合子基因盒。在 IntI1 阳性的 6 个分离物中发现了携带 1 类整合子基因盒的共轭质粒。在 CRPA 分离物中没有发现 2 类整合子。ERIC-PCR模式的树枝图分析表明,CRPA分离物之间没有克隆相似性,分离物没有通过交叉污染传播。结果发现,大多数有可能形成生物膜的 CRPA 分离物对碳青霉烯类以外的其他抗生素类具有高度耐药性,并能通过共轭耐药质粒共同转移某些耐药性(头孢他啶、头孢吡肟、环丙沙星、左氧氟沙星、哌拉西林-他唑巴坦)。这些菌株的耐药基因库有可能在临床上引起流行病,因此对这些菌株的耐药基因库进行分子流行病学跟踪会很有帮助。
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引用次数: 0
[Investigation of the Antibacterial Synergistic Activity of Fosfomycin-Teicoplanin Combination in Methicillin-Resistant Staphylococcus aureus Strains Isolated from Various Clinical Sample]. [福斯霉素-替考拉宁复方制剂对从不同临床样本中分离出的耐甲氧西林金黄色葡萄球菌菌株的抗菌协同作用研究]。
IF 1 4区 医学 Q4 MICROBIOLOGY Pub Date : 2024-04-01 DOI: 10.5578/mb.202498156
Rümeysa Tuba Biçer, Tuncer Özekinci, Mücahide Esra Koçoğlu

The aim of this study was to investigate the detection of teicoplanin and fosfomycin antibiotic susceptibility of methicillin-resistant Staphylococcus aureus (MRSA) strains by different methods and to evaluate the antibacterial synergistic effect of teicoplanin-fosfomycin combination by using checkerboard assay and time kill curve assay. Forty-five MRSA strains isolated from clinical samples in routine medical microbiology laboratory of Göztepe Prof. Dr. Süleyman Yalçın City Hospital were included in the study. In the first stage of the combination study, minimum inhibitory concentrations (MIC) were investigated by broth microdilution for teicoplanin and by both broth microdilution and agar dilution methods for fosfomycin. The combination of teicoplanin and fosfomycin was tested by the checkerboard method in 45 MRSA strains and combination effect was determined according to fractional inhibitory concentration index (ΣFIC) calculation. The synergistic effect and bactericidal activity of antibiotic combination were studied against a randomly selected strain from the strains used in the study by using time-kill method for 24 hours. As a result of teicoplanin and fosfomycin antibiotic susceptibility studies, all isolates were found to be susceptible to both antibiotics according to the susceptibility breakpoints determined by the European Committee on Antimicrobial Susceptibility Testing (EUCAST). A synergistic effect was found in 22 (49%), additive effect in 22 (49%) and indifferent effect in one (2%) of the 45 strains studied with the checkerboard method. The mean ΣFIC of 45 isolates was found to be 0.5. In the combination study of the antibiotics of the isolate that was studied with time-kill method, synergism was detected for 1/8 MIC concentrations at 12th hour and 24th hour and synergism at 1/4 MIC concentration at sixth hour, 12th hour and 24th hour. In the combination study of 1/4 MIC concentrations of antibiotics, bactericidal effect was detected at sixth hour and this effect was observed to disappear at 12th and 24th hours. High rate of synergistic antibacterial effect of teicoplanin-fosfomycin combination on MRSA isolates was demonstrated as a result of in vitro tests. Such studies conducted on antibiotic-resistant bacterial infections will provide clinicians different treatment options and will contribute to increasing survival. As a result of this study, provided that it is supported by future clinical studies, it can be stated that the teicoplanin-fosfomycin combination may be an effective treatment option in community and hospital-acquired infections caused by MRSA.

本研究旨在通过不同方法检测耐甲氧西林金黄色葡萄球菌(MRSA)菌株对替考拉宁和磷霉素的抗生素敏感性,并采用棋盘格检测法和时间杀伤曲线检测法评估替考拉宁-磷霉素复方制剂的抗菌增效作用。研究对象包括从 Göztepe Prof. Dr. Süleyman Yalçın 市医院常规医学微生物实验室的临床样本中分离出的 45 株 MRSA 菌株。在联合用药研究的第一阶段,采用肉汤微量稀释法检测了替考拉宁的最低抑菌浓度(MIC),采用肉汤微量稀释法和琼脂稀释法检测了磷霉素的最低抑菌浓度(MIC)。在 45 株 MRSA 菌株中采用棋盘格法检测了替考拉宁和磷霉素的复方制剂,并根据分数抑菌浓度指数(ΣFIC)计算确定了复方制剂的效果。采用时间杀灭法对研究中随机选取的菌株进行 24 小时杀菌,研究抗生素组合的协同效应和杀菌活性。根据欧洲抗菌药物敏感性测试委员会(EUCAST)确定的敏感性断点,对替考拉宁和磷霉素进行抗菌药物敏感性研究后发现,所有分离菌株都对这两种抗生素敏感。在使用棋盘格法研究的 45 株菌株中,有 22 株(49%)发现了协同效应,22 株(49%)发现了相加效应,1 株(2%)发现了无关效应。45 株分离菌株的平均ΣFIC 值为 0.5。在采用时间杀灭法对分离菌株进行的抗生素联合研究中,1/8 MIC 浓度的抗生素在第 12 小时和第 24 小时具有协同作用,1/4 MIC 浓度的抗生素在第 6 小时、第 12 小时和第 24 小时具有协同作用。在 1/4MIC浓度抗生素的联合研究中,第 6 小时检测到杀菌效果,第 12 和 24 小时这种效果消失。体外试验结果表明,替考拉宁-磷霉素复方制剂对 MRSA 分离物具有很高的协同抗菌效果。针对耐抗生素细菌感染开展的此类研究将为临床医生提供不同的治疗方案,并有助于提高存活率。通过这项研究,如果未来的临床研究支持这项研究,那么可以说替考拉宁-磷霉素复方制剂可能是治疗由 MRSA 引起的社区和医院感染的一种有效选择。
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引用次数: 0
[Determination of Capsule Types, Antibiotic Resistance Profiles and Phylogenetic Relationships of Group B Streptococcus Isolates Isolated from Patients Followed in Various Clinics and Polyclinics]. [确定从各诊所和综合医院随访患者中分离出的 B 组链球菌的胶囊类型、抗生素耐药性特征和系统发育关系]。
IF 1 4区 医学 Q4 MICROBIOLOGY Pub Date : 2024-01-01 DOI: 10.5578/mb.20249901
Suna Kızılyıldırım, Tülay Kandemir, Berfin Sucu, Fatih Köksal

Group B streptococci (GBS) are microorganisms that cause various systemic infections. In this study, it was aimed to investigate the capsule serotypes, antibiotic resistance and phylogenetic similarity relationship between GBS isolates. One hundred and ten GBS isolates isolated from female patients who admitted to Adana City Hospital with various complaints were included in the study. Kirby-Bauer disc diffusion method was used for the antibiotic resistance patterns and evaluated with CLSI criteria. The genes ermB, ermTR, mefA for erythromycin resistance and linB genes for clindamycin resistance were investigated by multiplex PCR method. Multiplex PCR method was used for GBS capsule serotyping. Similarity relationship between the isolates was analyzed by pulsed-field gel electrophoresis (PFGE) method. As a result of the study; all strains were found to be sensitive to penicillin and vancomycin. Erythromycin, clindamycin ofloxacin, and ceftriaxone resistance rates were observed as 60%, 11.8%, 6.4%, and 4.5%, respectively. The mefA gene was not found while 53% and 47% of the erythromycin-resistant isolates carried ermTR and ermB genes, respectively. The linB gene was not found in clindamycin-resistant GBS isolates. The capsule serotype distributions of GBSs were found as, Ib 42.7%, Ia 35.5%, III 10%, II 8.2%, and V 3.6%, respectively. In the analysis of the similarity relationships between GBS isolates with the PFGE method, no significant relationship was found. In conclusion, it was thought that more studies should be conducted to show the prevalence of GBS capsule serotypes and patterns of antibiotic resistance.

乙型链球菌(GBS)是一种可引起各种全身感染的微生物。本研究旨在调查 GBS 分离物的胶囊血清型、抗生素耐药性和系统发育相似性关系。研究对象包括从阿达纳市医院因各种主诉入院的女性患者中分离出的 110 株 GBS 分离物。采用柯比鲍尔(Kirby-Bauer)盘扩散法检测抗生素耐药性模式,并根据 CLSI 标准进行评估。采用多重 PCR 法检测了红霉素耐药基因 ermB、ermTR 和 mefA,以及克林霉素耐药基因 linB。多重 PCR 法用于 GBS 胶囊血清分型。采用脉冲场凝胶电泳(PFGE)方法分析了分离物之间的相似性关系。研究发现,所有菌株都对青霉素和万古霉素敏感。红霉素、林可霉素、氧氟沙星和头孢曲松的耐药率分别为 60%、11.8%、6.4% 和 4.5%。未发现 mefA 基因,而 53% 和 47% 的红霉素耐药分离物分别带有 ermTR 和 ermB 基因。耐克林霉素的 GBS 分离物中未发现 linB 基因。发现 GBS 的胶囊血清型分布分别为:Ib 42.7%、Ia 35.5%、III 10%、II 8.2%和 V 3.6%。用 PFGE 法分析 GBS 分离物之间的相似性关系时,没有发现明显的关系。总之,人们认为应开展更多的研究,以显示 GBS 胶囊血清型的流行情况和抗生素耐药性模式。
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引用次数: 0
[Investigation of the Inhibition Effect of Acanthamoeba Cell-Free Supernatants Against Pseudomonas aeruginosa]. [无阿卡胞浆对铜绿假单胞菌抑制作用的研究]。
IF 1 4区 医学 Q4 MICROBIOLOGY Pub Date : 2024-01-01 DOI: 10.5578/mb.20249950
Çise Kebabcı, Zuhal Zeybek

Free-living amoebae belonging to the genus Acanthamoeba are microorganisms that live in air, soil and aquatic environments. In humans, they cause infections such as amoebic keratitis, graulamotous amoebic encephalitis that are difficult to treat and can be fatal. In addition, it is known that they contribute to the replication of bacteria and increase their pathogenicity by being a host for various bacteria. However, information on its inhibitory properties against bacteria and its production of antimicrobial agents is very limited. In this context, in this study, it was aimed to investigate whether cell-free supernatants of Acanthamoeba strains have antibacterial effects against Pseudomonas aeruginosa isolates. Four different Acanthamoeba strains (A10, A13, A14, U.GÖL) isolated from aquatic environments in our country were selected and used in the study, P.aeruginosa isolates (PA2, PA3, PA4, PA5) were selected from clinical strains belonging to patients in our country. Acanthamoeba castellanii ATCC 50373 and P.aeruginosa ATCC 27853 were used as standard strains. P.aeruginosa isolates were grown on nutrient agar at 37 °C and Acanthamoeba strains were grown on E.coli spread non-nutrient agar at 30 °C under aerobic conditions. Pepton yeast extract glucose (PYG) medium supplemented with penicillin and streptomycin was used to obtain axenic cultures of Acanthamoeba strains. After the centrifugation of axenic cultures at 3000 rpm for five minutes, Acanthamoeba-cell-free supernatants were obtained by filtering the supernatant part through a sterile filter with a pore diameter of 0.22 µm. The antibacterial activities of these supernatants against P.aeruginosa isolates were determined using the colony counting method. Analysis of each Acanthamoeba-cell-free supernatants was performed according to the GC-MS method. Acanthamoeba-cell-free supernatants were found to have varying degrees of inhibitory effects (3.9-91.5%) against tested P.aeruginosa isolates. It was determined that the cell-free supernatant of A.castellanii ATCC 50373 strain showed the highest antibacterial effect (91.5%) against PA5 isolate. A14 strain showed similar inhibitory effects (89.4%) against the same Pseudomonas isolate. The average inhibitory effect of most of the Acanthamoeba strains of our country was found to be higher than that of the reference strain A.castellanii ATCC 50492. It is thought that the compounds responsible for the anti-Pseudomonas activity of the tested Acanthamoeba strains may be fructose, phosphoric acid, galactose, N-Acetylphenylalanine and glucopyranose determined as major compounds. This is the first study showing the anti-Pseudomonas activity of microorganisms of the genus Acanthamoeba living in the waters of our country. Acanthamoeba, which is widely found in nature, appears to be a good source for new antimicrobial agents.

属于阿卡阿米巴属的自由生活阿米巴是一种生活在空气、土壤和水生环境中的微生物。在人体内,它们会引起阿米巴角膜炎、葡萄胎阿米巴脑炎等感染,这些感染难以治疗,甚至可能致命。此外,人们还知道它们有助于细菌的复制,并通过成为各种细菌的宿主而增加其致病性。然而,有关它对细菌的抑制特性及其生产抗菌剂的信息却非常有限。在这种情况下,本研究旨在探讨棘阿米巴菌株的无细胞上清液是否对铜绿假单胞菌分离物具有抗菌作用。本研究选取了从我国水生环境中分离的四种不同的棘阿米巴菌株(A10、A13、A14、U.GÖL),铜绿假单胞菌分离物(PA2、PA3、PA4、PA5)选自我国患者的临床菌株。卡氏棘阿米巴菌 ATCC 50373 和绿脓杆菌 ATCC 27853 被用作标准菌株。铜绿假单胞菌分离株在 37 ℃ 营养琼脂上生长,而棘阿米巴菌株则在 30 ℃ 有氧条件下在大肠杆菌扩散的非营养琼脂上生长。使用添加青霉素和链霉素的百普顿酵母提取物葡萄糖(PYG)培养基获得棘阿米巴菌株的轴突培养物。将轴向培养物以 3000 rpm 的转速离心 5 分钟后,用孔径为 0.22 µm 的无菌过滤器过滤上清液,得到不含棘阿米巴细胞的上清液。采用菌落计数法测定这些上清液对铜绿假单胞菌分离株的抗菌活性。根据气相色谱-质谱(GC-MS)方法对每种不含棘阿米巴细胞的上清液进行了分析。结果发现,无阿卡胞上清液对铜绿假单胞菌有不同程度的抑制作用(3.9%-91.5%)。结果表明,A.castellanii ATCC 50373 菌株的无细胞上清液对 PA5 分离物的抗菌效果最高(91.5%)。A14 菌株对同一假单胞菌分离物也有类似的抑菌效果(89.4%)。发现我国大多数棘阿米巴菌株的平均抑制效果高于参考菌株 A.castellanii ATCC 50492。据此推测,被测棘阿米巴菌株具有抗假单胞菌活性的主要化合物可能是果糖、磷酸、半乳糖、N-乙酰苯丙氨酸和葡萄糖。这是首次显示生活在我国水域中的棘阿米巴属微生物具有抗假单胞菌活性的研究。广泛存在于自然界中的棘阿米巴似乎是新抗菌剂的良好来源。
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引用次数: 0
[Investigation of the Relationship Between Akkermansia Genomic Diversity in Gut Microbiota and Parkinson's Disease Dementia]. [调查肠道微生物群中 Akkermansia 基因组多样性与帕金森病痴呆症之间的关系]。
IF 1 4区 医学 Q4 MICROBIOLOGY Pub Date : 2024-01-01 DOI: 10.5578/mb.20249951
Muzaffer Arıkan, Tuğçe Kahraman Demir, Zeynep Yıldız, Nesrin Helvacı Yılmaz, Aysu Şen, Lütfü Hanoğlu, Süleyman Yıldırım

Although it is known that the relative abundance of Akkermansia, a bacterial genus commonly associated with health, increases in the gut microbiota of Parkinson's disease (PD) patients, the exact reason for this increase remains unclear. This study was aimed to identify potential changes in Akkermansia within the gut microbiota of PD patients in Türkiye. For this purpose, shotgun metagenomics and a novel Akkermansia genus-specific amplicon sequencing technique was used to investigate the presence of specific Akkermansia strains associated with cognitive impairment (CI) stages in PD and to examine potential genes within these strains. In this context, four gut microbiota samples from Türkiye -three PD with dementia (PDD) and one healthy control without CI (HC)- were analyzed by shotgun metagenomics and metagenome-assembled genomes assigned to Akkermansia genus were reconstructed. Then, a custom database was created by combining these genomes with the Akkermansia genomes in public databases and next generation sequencing (NGS) compatible primers specific to the genus Akkermansia were designed using this database. After optimization of amplification and library preparation steps for genus-specific next generation sequencing, gut microbiota samples from 64 PD patients [32 PDD and 32 PD with mild CI (PD-MCI)] and 26 HCs were analyzed by genus-specific amplicon sequencing. The results revealed the presence of seven strains assigned to Akkermansia muciniphila in gut microbiota samples, two of which showed significant distribution differences (p< 0.05) between demented (PDD) and non-demented groups (PD-MCI, HC). When gene contents of the detected Akkermansia genomes were examined through comparative genomic analysis, the presence of 12 genes only in Akkermansia genomes specific to non-demented groups were predicted. The annotations of these genes showed that they were not reported before with unknown functions. In this study, for the first time, gut microbiota samples from PD patients in Türkiye were analyzed using shotgun metagenomics, a novel genus-specific amplicon sequencing method was developed specifically for the analysis of Akkermansia genus, and then Akkermansia strains and genes potentially associated with CI stages in PD were identified using this method. The results underscore that investigating the species or strain level differences could help better understanding of the changes associated with PD in the human gut microbiota.

众所周知,在帕金森病(PD)患者的肠道微生物群中,通常与健康有关的细菌属 Akkermansia 的相对丰度会增加,但这种增加的确切原因仍不清楚。本研究旨在确定土耳其帕金森病患者肠道微生物群中 Akkermansia 的潜在变化。为此,研究人员采用了霰弹枪元基因组学和新型 Akkermansia 属特异性扩增子测序技术来研究与帕金森病认知障碍(CI)阶段相关的特定 Akkermansia 菌株的存在情况,并检测这些菌株中的潜在基因。在此背景下,通过霰弹枪元基因组学分析了来自土耳其的四个肠道微生物群样本--三个痴呆症患者(PDD)和一个无 CI 的健康对照(HC)--并重建了分配给 Akkermansia 属的元基因组。然后,将这些基因组与公共数据库中的 Akkermansia 基因组相结合,创建了一个定制数据库,并利用该数据库设计了 Akkermansia 属的下一代测序(NGS)兼容引物。在优化了扩增和文库制备步骤以进行属特异性下一代测序后,通过属特异性扩增片段测序分析了来自 64 名 PD 患者(32 名 PDD 和 32 名 PD 伴轻度 CI(PD-MCI))和 26 名 HC 的肠道微生物群样本。结果显示,肠道微生物群样本中存在 7 株属于 Akkermansia muciniphila 的菌株,其中 2 株在痴呆组(PDD)和非痴呆组(PD-MCI、HC)之间存在显著的分布差异(p< 0.05)。通过比较基因组分析对检测到的Akkermansia基因组中的基因内容进行研究,发现12个基因仅存在于非痴呆组的Akkermansia基因组中。这些基因的注释表明,它们的功能未知,以前从未被报道过。本研究首次使用霰弹枪元基因组学方法分析了土耳其帕金森病患者的肠道微生物群样本,开发了一种专门用于分析 Akkermansia 属的新型属特异性扩增片段测序方法,然后使用该方法鉴定了可能与帕金森病 CI 阶段相关的 Akkermansia 菌株和基因。研究结果表明,研究物种或菌株水平的差异有助于更好地了解人类肠道微生物群与腹泻相关的变化。
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引用次数: 0
[Distribution of the Prevalence of Human Leukocyte Antigen (HLA)-B*57:01 Positivity in HIV-1 Infected Individuals and Its Effects on Treatment: Türkiye Map-Buhasder Working Group]. [人类白细胞抗原(HLA)-B*57:01 阳性在 HIV-1 感染者中的分布及其对治疗的影响:图尔基耶 Map-Buhasder 工作组]。
IF 1 4区 医学 Q4 MICROBIOLOGY Pub Date : 2024-01-01 DOI: 10.5578/mb.20249903
Seyit Ali Büyüktuna, Caner Öksüz, Alper Tahmaz, Figen Sarıgül Yıldırım, Melda Türken, Özgür Günal, Şeyma Topal, Ali İrfan Baran, Burak Sarıkaya, Semiha Çelik Ekinci, Selçuk Kaya, Sevil Alkan Çeviker, Adalet Aypak, Pınar Yürük Atasoy, Dilara İnan, Adem Köse, Nevind Koç İnce, Seniha Şenbayrak, Şafak Kaya, Müge Özgüler, Emine Kübra Dindar Demiray, Şükran Köse
<p><p>Human immunodeficiency virus (HIV)/acquired immundeficiency syndrome (AIDS) is a critical global public health problem that significantly affects both life expectancy and the overall quality of life of individuals in all age groups. The landscape of HIV infection has changed significantly in recent years due to the introduction of effective combination antiretroviral therapies (ART). A key component of first-line ART regimens for HIV treatment is abacavir, a nucleoside HIV reverse transcriptase inhibitor. Although abacavir is effective in suppressing viral replication and managing disease, its clinical utility is overshadowed by the potential for life-threatening hypersensitivity reactions in HLA-B*57:01-positive patients. In our country, local data obtained from various centers regarding the prevalence of HLA-B*57:01 in HIV-1-infected patients are available. In this study, it was aimed to determine the prevalence of the HLA-B*57:01 genotype in HIV-infected patients who were followed up and treated in many regions of our country. This retrospective study consists of the data of the patients aged 18 years and over diagnosed with HIV-1 infection between 01.01.2019 and 31.07.2022. Age, gender, place of birth, mode of transmission of the disease, death status, CD4+ T cell count and HIV RNA levels at the first clinical presentation, HLA-B*57:01 positivity, and the method used, clinical stage of the disease, virological response time with the treatment they received were recorded from the patient files. Data were collected from 16 centers and each center used different methods to detect HLA-B*57:01. These methods were sequence-specific oligonucleotide probe hybridization (SSOP), DNA sequence-based typing (SBT), single-specific primer-polymerase chain reaction (SSP-PCR), allele-specific PCR (AS-PCR) and quantitative PCR (Q-PCR). A total of 608 HIV-infected individuals, 523 males (86%) and 85 females (14%), were included in the study. The mean age of the patients was 36.9 ± 11.9 (18-73) years. The prevalence of HLA-B*57:01 allele was found to be 3.6% (22 patients). The number of CD4+ T lymphocytes in HLA-B*57:01 allele-positive patients was > 500/ mm3 in 10 patients (45.5%), while the number of CD4+ T lymphocytes in HLA-B*57:01 negative patients was > 500/mm3 in 216 patients (36.9%) (p> 0.05). Viral load at the time of diagnosis was found to be lower in patients with positive HLA-B*57:01 allele but it was not statistically significant (p> 0.05). Although different treatment algorithms were used in the centers following the patients, it was observed that the duration of virological response was shorter in HLA-B*57:01 positive patients (p= 0.006). Although the presence of the HLA-B*57:01 allele has a negative impact due to its association with hypersensitivity, it is likely to continue to attract interest due to its association with slower progression of HIV infection and reduced risk of developing AIDS. In addition, although the answer to the questi
人体免疫缺陷病毒(HIV)/获得性免疫缺陷综合症(AIDS)是一个严重的全球性公共卫生问题,严重影响着各年龄段人群的预期寿命和整体生活质量。近年来,由于引入了有效的抗逆转录病毒联合疗法(ART),HIV 感染的情况发生了显著变化。阿巴卡韦是核苷类艾滋病逆转录酶抑制剂,是一线抗逆转录病毒疗法治疗艾滋病的关键组成部分。虽然阿巴卡韦在抑制病毒复制和控制病情方面效果显著,但其在 HLA-B*57:01 阳性患者中可能引发危及生命的超敏反应,这给阿巴卡韦的临床应用蒙上了阴影。在我国,有来自不同中心的关于 HLA-B*57:01 在 HIV-1 感染者中流行情况的本地数据。本研究旨在确定在我国多个地区接受随访和治疗的 HIV 感染者中 HLA-B*57:01 基因型的流行情况。这项回顾性研究包括 2019 年 1 月 1 日至 2022 年 7 月 31 日期间被诊断为感染 HIV-1 的 18 岁及以上患者的数据。从患者档案中记录了他们的年龄、性别、出生地、疾病传播方式、死亡状况、首次临床表现时的 CD4+ T 细胞计数和 HIV RNA 水平、HLA-B*57:01 阳性、使用的方法、疾病的临床阶段、接受治疗后的病毒学应答时间。数据来自 16 个中心,每个中心使用不同的方法检测 HLA-B*57:01。这些方法包括序列特异性寡核苷酸探针杂交(SSOP)、DNA序列分型(SBT)、单特异性引物聚合酶链反应(SSP-PCR)、等位基因特异性 PCR(AS-PCR)和定量 PCR(Q-PCR)。研究共纳入了 608 名艾滋病毒感染者,其中男性 523 人(占 86%),女性 85 人(占 14%)。患者的平均年龄为 36.9 ± 11.9 (18-73) 岁。HLA-B*57:01等位基因的患病率为3.6%(22名患者)。HLA-B*57:01等位基因阳性患者中CD4+ T淋巴细胞数量大于500/mm3的有10例(45.5%),而HLA-B*57:01阴性患者中CD4+ T淋巴细胞数量大于500/mm3的有216例(36.9%)(P> 0.05)。HLA-B*57:01等位基因阳性患者诊断时的病毒载量较低,但无统计学意义(P> 0.05)。虽然跟踪患者的中心采用了不同的治疗算法,但据观察,HLA-B*57:01 阳性患者的病毒学应答持续时间较短(p= 0.006)。尽管 HLA-B*57:01 等位基因的存在因其与高过敏性有关而产生负面影响,但因其与减缓艾滋病病毒感染进展和降低发展为艾滋病的风险有关,可能会继续引起人们的关注。此外,尽管人们正在寻找在开始使用含阿巴卡韦的抗逆转录病毒疗法治疗艾滋病病毒感染之前对患者进行 HLA-B*57:01 筛查是否具有成本效益的答案,但艾滋病病毒治疗指南似乎将继续建议进行筛查,以确定有这方面风险的患者。
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引用次数: 0
[Investigation of the Effect of Farnesol on Biofilm Formation by Candida albicans and Candida parapsilosis Complex Isolates]. [法尼醇对白色念珠菌和副丝状念珠菌复合菌株生物膜形成的影响研究]。
IF 1 4区 医学 Q4 MICROBIOLOGY Pub Date : 2024-01-01 DOI: 10.5578/mb.20249905r
Berna Erdal, Bensu Baylan, Bahadır Batar, Ali Öztürk, Birol Topçu
<p><p>The incidence of infections caused by Candida species has significantly increased over the past three decades. Candida albicans is commonly recognized as the primary causative agent in cases of candidiasis; however, non-albicans Candida species, including Candida parapsilosis, are also frequently defined as pathogens. Treatment-resistant infections arise as a result of biofilm formation, which is one of the effective mechanisms in the pathogenesis of Candida infections. However, the mechanisms of action of farnesol, a quorum sensing (QS) system molecule, on biofilm formation by Candida species remain unclear. This study aimed to demonstrate the changes in the biofilm biomass of C.albicans and C.parapsilosis complex isolates in the presence of farnesol and reveal the expression of the EFG1 and BCR1 genes, which are believed to play a role in the production of QS molecules, using quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) analysis. C.albicans (n= 91) and C.parapsilosis complex (n= 29) isolates obtained from different clinical samples were included in the study. The minimum inhibitory concentration (MIC) values of farnesol were determined using the broth microdilution method according to the M27-A3 protocol of the Clinical and Laboratory Standards Institute (CLSI). The biofilm biomass of the isolates was examined without farnesol and at the MIC-0 and MIC-2 concentrations of farnesol. Changes in the expression of the biofilm-associated EFG1 and BCR1 genes were investigated using qRT-PCR. According to the results of the study, the MIC values of farnesol were detected in the range of 1-2 mM in 82.4% (n= 75) of the C.albicans isolates and in the range of 0.5-1 mM in 72.4% (n= 21) of the C.parapsilosis complex isolates. Of the C.albicans isolates, 27 (29.7%) exhibited a strong biofilm formation and 58 (63.7%) demonstrated a weaker biofilm formation, while these rates were 34.4% (n= 10) and 62.1% (n= 18), respectively, for the C.parapsilosis complex isolates. At the MIC-0 and MIC-2 concentrations, farnesol was observed to reduce biofilm biomass among C.albicans (n= 24, 88.9%) and C.parapsilosis complex (n= 8, 80.0%) isolates that formed strong biofilms and observed to increase biofilm biomass among those that formed weak biofilms [60.3% (n= 35) and 55.6% (n= 10), respectively]. On completion of the qRT-PCR analysis supporting the results of the biofilm experiment, it was determined that the expressions of the EFG1 and BCR1 genes decreased at the MIC-0 and MIC-2 concentrations of farnesol among the strong biofilm-forming C.albicans and C.parapsilosis complex isolates, but there was an increase in gene expressions among the weak biofilm-forming isolates. In addition to the antifungal effect of farnesol on Candida species, this study provided data on the efficacy of the MIC-0 and MIC-2 concentrations of farnesol against Candida biofilm biomass. Although our results suggest that farnesol can be used as an alternative agent to
过去 30 年来,念珠菌感染的发病率大幅上升。白色念珠菌通常被认为是念珠菌病的主要致病菌,但包括副丝状念珠菌在内的非白色念珠菌也经常被定义为病原体。耐药性感染是生物膜形成的结果,而生物膜是念珠菌感染致病的有效机制之一。然而,法定量传感(QS)系统分子法尼醇对念珠菌生物膜形成的作用机制仍不清楚。本研究旨在利用反转录酶聚合酶链式反应(qRT-PCR)定量分析法尼醇存在时白念珠菌和副丝状念珠菌复合体生物膜生物量的变化,并揭示 EFG1 和 BCR1 基因的表达,这些基因被认为在 QS 分子的产生中发挥作用。从不同临床样本中分离出的白僵菌(n= 91)和副丝虫(n= 29)被纳入研究。法尼醇的最小抑菌浓度(MIC)值是根据临床和实验室标准协会(CLSI)的 M27-A3 协议,采用肉汤微稀释法测定的。在没有法尼醇的情况下,以及在法尼醇的 MIC-0 和 MIC-2 浓度下,对分离菌株的生物膜生物量进行了检测。使用 qRT-PCR 技术研究了生物膜相关 EFG1 和 BCR1 基因的表达变化。研究结果表明,82.4%(n= 75)的白僵菌分离物的法尼醇 MIC 值范围为 1-2 mM,72.4%(n= 21)的白僵菌复合体分离物的法尼醇 MIC 值范围为 0.5-1 mM。在白僵菌分离物中,27 个(29.7%)表现出较强的生物膜形成,58 个(63.7%)表现出较弱的生物膜形成,而副丝状菌复合体分离物的生物膜形成率分别为 34.4%(10 个)和 62.1%(18 个)。在 MIC-0 和 MIC-2 浓度下,观察到法尼醇降低了形成强生物膜的白僵菌(样本数=24,88.9%)和副丝状菌复合体(样本数=8,80.0%)分离物的生物膜生物量,增加了形成弱生物膜的分离物的生物膜生物量[分别为 60.3%(样本数=35)和 55.6%(样本数=10)]。在完成支持生物膜实验结果的 qRT-PCR 分析后,确定在法尼醇的 MIC-0 和 MIC-2 浓度下,形成强生物膜的白僵菌和副丝状菌复合体分离物的 EFG1 和 BCR1 基因表达量减少,但形成弱生物膜的分离物的基因表达量增加。除了法尼醇对白色念珠菌的抗真菌作用外,本研究还提供了 MIC-0 和 MIC-2 浓度的法尼醇对白色念珠菌生物膜生物量的功效数据。尽管我们的研究结果表明,法尼醇可作为一种替代药剂来减少念珠菌感染中生物膜的形成,但还需要进一步的研究来支持。此外,这项研究还首次确定了在法尼醇存在的情况下,副丝状菌复合体分离物中 EFG1 和 BCR1 基因的表达情况,因此具有重要意义。
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引用次数: 0
[A Case Report of Monkeypox Disease in an Adolescent]. [青少年猴痘病例报告]。
IF 1 4区 医学 Q4 MICROBIOLOGY Pub Date : 2024-01-01 DOI: 10.5578/mb.20249909
Leyla Beşel, Önder Kılıçaslan, Çiğdem Kırmacı, Didem Kızmaz İsançlı, Irmak Emre, Adem Karbuz

Monkeypox virus (MPXV) infection is a zoonotic disease characterized by smallpox-like rashes. It is endemic in Central and West Africa. The World Health Organization (WHO) declared the disease as an epidemic due to a significant increase in the number of reported cases, starting from Europe and spreading to other regions, particularly in certain areas, in May 2022. On July 23, 2022, it was recognized as a public health problem of international importance. Our country has also been affected by this epidemic, and the official number of reported cases is twelve. In this case report, an adolescent case diagnosed with MPXV infection was presented. A 17-year-old male patient admitted with the complaints of sores around the mouth and genital area, fever and headache. The patient had a history of sexual contact with three different males in the last six months. Honey-colored crusted papules and plaques were observed in the perioral area, as well as crusted papules on the penile and gluteal areas. Ulcerative sores were present in the oral cavity. Laboratory tests for sexually transmitted diseases confirmed the patient's HIV-positive status and MPXV infection through PCR (polymerase chain reaction) testing. Antiviral treatment for human immunodeficiency virus (HIV) was initiated after the HIV RNA level resulted in 263000 copies/mL. Additionally, a glycopeptide was added to the treatment regimen when methicillin-resistant Staphylococcus aureus growth was detected in the swab culture taken from the wounds on the patient's face. No specific treatment was administered for MPXV due to the patient's uncomplicated clinical course and overall well-being. This case report aims to raise awareness about monkeypox disease in children by highlighting the clinical findings and potential risk factors.

猴痘病毒(MPXV)感染是一种以类似天花的皮疹为特征的人畜共患疾病。它在非洲中部和西部流行。2022 年 5 月,由于报告病例数量显著增加,世界卫生组织(WHO)宣布该疾病为流行病。2022 年 7 月 23 日,该疾病被确认为具有国际重要性的公共卫生问题。我国也受到了这一流行病的影响,官方报告病例数为 12 例。本病例报告介绍了一例确诊感染 MPXV 的青少年病例。一名 17 岁的男性患者入院时主诉口腔和生殖器周围有溃疡、发烧和头痛。患者在过去 6 个月中曾与 3 名不同的男性有性接触史。口周出现蜜色结痂丘疹和斑块,阴茎和臀部也有结痂丘疹。口腔出现溃疡。通过聚合酶链反应(PCR)检测,性传播疾病实验室检测证实患者为艾滋病毒阳性和 MPXV 感染。在艾滋病毒 RNA 水平达到 263000 拷贝/毫升后,患者开始接受人类免疫缺陷病毒(HIV)抗病毒治疗。此外,当从患者面部伤口采集的拭子培养液中检测到耐甲氧西林金黄色葡萄球菌生长时,治疗方案中添加了糖肽。由于患者的临床病程并不复杂,整体状况良好,因此没有对 MPXV 进行特殊治疗。本病例报告旨在通过强调临床发现和潜在风险因素,提高人们对儿童猴痘疾病的认识。
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引用次数: 0
[Investigation of Cryptococcus Colonization and Mating Genotype in Environmental Samples]. [环境样本中隐球菌的定植和交配基因型调查]。
IF 1 4区 医学 Q4 MICROBIOLOGY Pub Date : 2024-01-01 DOI: 10.5578/mb.20249904
Deniz Alkaya, Engin Kaplan, Çağrı Ergin, Macit İlkit, Aylin Döğen

Cryptococcus species are fungal pathogens that pose a serious threat to human life and can cause meningoencephalitis in immunocompromised and healthy individuals. It was estimated that approximately 112000 people die every year due to cryptococcal-related infections all over the world, especially in immunocompromised individuals. Cryptococcus species can be found in soil, bat dung, pigeon droppings, and various tree species in addition to humans. Despite the majority of Cryptococcus species being haploid opportunistic human pathogens, it is known that the ability to undergo sexual reproduction plays a significant role in the expansion of species distribution and the increase in virulence. In Cryptococcus species, sexual reproduction is governed by the mating genotype gene region called the MAT locus. Pathogenic Cryptococcus species have two mating types (MATa and MATα), defined by the presence of one of two alternative alleles at a single MAT locus. In this study, various tree species (eucalyptus, olive and carob) in a total of seven regions in Mersin (Gülnar, Göksu, Narlıkuyu, Ayaş, Kızkalesi, and Tarsus) and Hatay provinces were examined to detect Cryptococcus species. The aim of this study was to determine the environmental distribution and sexual genotypes of Cryptococcus species in these regions. In the present study, samples were collected from a total of 750 trees, including olive, eucalyptus, and carob trees. The samples were incubated on Staib agar medium containing 0.1% biphenyl and 0.5% chloramphenicol. Colonies that formed brown pigment were identified as C.neoformans using conventional and molecular methods. The sexual genotypes were determined by comparing the lengths of the STE20 gene from the isolates compared with those of reference C.neoformans strains. Growth was observed in 97 (12.9%) of 750 samples collected from eucalyptus (n= 236), olive (n= 303) and carob (n= 211) trees. All 97 isolates were determined to be C.neoformans var. grubii. The highest positivity was found in Narlıkuyu (78.2%), and from carob (9.4%) and olive (3.5%) trees. Cryptococcus species was not detected in any of the samples derived from eucalyptus trees. Based on the lengths of the STE20 gene, it was determined that all C.neoformans var. grubii isolates were in the MAT Aα genotype. The data obtained regarding the environmental distribution of Cryptococcus species and the distribution of genes involved in sexual reproduction are believed to provide valuable guidance in terms of the potential clinical implications of environmental Cryptococcus hotspots and regional species characteristics in our country.

隐球菌是对人类生命构成严重威胁的真菌病原体,可导致免疫力低下者和健康人患脑膜脑炎。据估计,全世界每年约有 11 2 000 人死于隐球菌相关感染,尤其是免疫力低下者。除人类外,隐球菌还存在于土壤、蝙蝠粪便、鸽子粪便和各种树种中。尽管大多数隐球菌都是单倍体的机会性人类病原体,但众所周知,有性生殖能力在扩大物种分布和增强毒力方面发挥着重要作用。在隐球菌中,有性生殖受称为 MAT 基因座的交配基因型基因区域控制。致病性隐球菌有两种交配类型(MATa 和 MATα),由单个 MAT 基因座上存在的两个备选等位基因之一决定。本研究对梅尔辛省(Gülnar、Göksu、Narlıkuyu、Ayaş、Kızkalesi 和 Tarsus)和哈塔伊省共七个地区的各种树种(桉树、橄榄树和角豆树)进行了检测,以发现隐球菌。本研究的目的是确定这些地区隐球菌的环境分布和有性基因型。本研究共采集了 750 棵树的样本,包括橄榄树、桉树和角豆树。样本在含有 0.1% 联苯和 0.5% 氯霉素的 Staib 琼脂培养基上培养。用传统方法和分子方法鉴定形成棕色色素的菌落为新霉菌。通过比较分离菌株与参考 C.neoformans 菌株的 STE20 基因长度,确定性基因型。在从桉树(n= 236)、橄榄树(n= 303)和角豆树(n= 211)采集的 750 份样本中,有 97 份(12.9%)观察到了生长。所有 97 个分离物均被确定为 C.neoformans var.阳性率最高的是 Narlıkuyu(78.2%)、角豆树(9.4%)和橄榄树(3.5%)。桉树样本中均未检测到隐球菌。根据 STE20 基因的长度,可以确定所有 C.neoformans 变种 grubii 分离物都属于 MAT Aα 基因型。所获得的有关隐球菌物种环境分布和有性生殖基因分布的数据,相信能为我国环境隐球菌热点地区的潜在临床影响和区域物种特征提供有价值的指导。
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